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Objectives: Long-acting bronchodilators are important treatments for chronic obstructive pulmonary disease (COPD) and adequate medication adherence decreases COPD exacerbations, especially in reducing the hazard of influenza infection. Therefore, the study aim was to evaluate adherence of long-acting bronchodilator treatment and the risk of influenza in patients with COPD. Methods: This retrospective nested case-control study included patients with newly diagnosed COPD from 2012 to 2018. Cases with influenza infection were defined and matched to 2 randomly selected controls. The influenza infection date was the index date. Conditional logistic regressions were used to estimate odds ratios of influenza from proportion of days covered (PDC) of long-acting bronchodilators measured in one year before the index date. Adherence was divided into high adherence (PDC ≥80 %) and low adherence (PDC

Background: We investigated the impacts of the standard treatment durations of and adherence to standard anti-tuberculous therapy (ATT) on recurrence after the successful completion of tuberculosis (TB) treatment. Methods: We recruited patients with TB who had received treatment for six or nine months from the 2008–2017 databases of the Taiwanese National Health Insurance Research Database. Treatment duration and adherence to standard ATT were analyzed for their impacts on recurrence within two years. Complete adherence to standard ATT was defined as daily use of ethambutol, isoniazid, pyrazinamide, and rifampin for the first two months, and daily use of isoniazid and rifampin for the first six months. Results: A total of 33,298 TB patients with new-onset TB were identified and classified into two groups by treatment duration: six months (n = 25,849, 77.63%) and nine months (n = 7449). Sex and age distributions varied between the groups. Treatment duration did not affect TB recurrence within two years (adjusted hazard ratio (AHR): 1.18, 95% confidence interval (C.I.) [0.96–1.44], p = 0.1156). Multivariable logistic regression showed that incomplete adherence to standard anti-tuberculous therapy (80–89% and 90–99% standard anti-TB therapy, AHR: 1.57, 95% C.I. [1.26–1.95], and 1.63, 95% C.I. [1.26–2.06], respectively, p

Background: Type 2 diabetes mellitus (DM) is a risk factor for mycobacterial pulmonary infections (MPI), including tuberculosis (TB) and nontuberculous mycobacterial lung disease (NTM-LD). Dipeptidyl peptidase IV inhibitor (DPP4i), a common DM medication, has an immune-modulation effect that raises concerns about developing MPI. However, there is scarce research on the topic. Methods: This retrospective study was conducted in a tertiary-referral center in Taiwan from 2009 to 2016. Patients with type 2 DM who were receiving any DM medication were enrolled. TB and NTM-LD were defined by microbiological criteria. We analyzed the risk of MPI in DPP4i users using Cox proportional hazard regression with adjusted inverse probability of treatment weighting. Results: A total of 9963 patients were included. Among them, 3931 were classified as DPP4i users, and 6032 patients were DPP4i nonusers. DPP4i users had no increase in incidences of MPI (604 vs. 768 per 100,000 person-years, p = 0.776), NTM-LD (174 vs. 255 per 100,000 person-years, p = 0.228), and TB (542 vs. 449 per 100,000 person-years, p = 0.663) relative to those of DPP4i nonusers. After adjustment, the adjusted hazard ratios for MPI (aHR: 1.07, 95% CI: 0.79–1.45), TB (aHR: 1.15, 95% CI: 0.81–1.64) and NTM-LD (aHR: 0.85, 95% CI: 0.49–1.47) were not significantly increased relative to those of nonusers. The subgroup analysis also showed that DPP4i use did not increase the risk of MPI in different DM severities and comorbidities. Conclusions: According to our large cohort study, DPP4i use is safe for patients with type 2 DM and might not increase the risk of MPI.

Summary: In this research, we aimed to harness machine learning to predict the imminent risk of acute exacerbation in chronic obstructive pulmonary disease (AECOPD) patients. Utilizing retrospective data from electronic medical records of two Taiwanese hospitals, we identified 26 critical features. To predict 3- and 6-month AECOPD occurrences, we deployed five distinct machine learning algorithms alongside ensemble learning. The 3-month risk prediction was best realized by the XGBoost model, achieving an AUC of 0.795, whereas the XGBoost was superior for the 6-month prediction with an AUC of 0.813. We conducted an explainability analysis and found that the episode of AECOPD, mMRC score, CAT score, respiratory rate, and the use of inhaled corticosteroids were the most impactful features. Notably, our approach surpassed predictions that relied solely on CAT or mMRC scores. Accordingly, we designed an interactive prediction system that provides physicians with a practical tool to predict near-term AECOPD risk in outpatients.

Background: Lung cancer patients are high-risk for active tuberculosis (TB); however, fragility and drug-drug interaction might lead to TB treatment interruption. TB treatment incompletion occurrence and predictors among lung cancer patients remain unclear. Methods: We recruited lung cancer patients with new-onset TB from Taiwan Cancer Registry and Taiwanese National Health Insurance 2007–2015 databases. TB treatment incompletion was the identified primary outcome, and associated risk factors were analyzed. Results: A total of 1155 lung cancer patients with new-onset TB were identified and classified as treatment incompletion (n=706, 61.13%) or completion (n=449). Gender and age distribution was similar in both groups. Under multivariable logistic regression, advanced cancer (stage III and IV) and no first-line TB drugs use were independent factors for treatment incompletion; but older age was not significant. For patients surviving >1 year since TB diagnosis, independent factors for treatment incompletion included no first-line TB drugs use (except pyrazinamide) and absence of hypertension. Cancer stage had borderline significance. Conclusions: TB treatment incompletion occurred in 61.13% of lung cancer patients. Clinicians should carefully titrate anti-TB medications and monitor side effects in lung cancer patients, especially those with treatment incompletion risk factors, to avoid treatment interruption due to fragility and/or drug intolerance.

Seismic responses of the Zipingpu concrete face rockfill dam were analyzed using the finite element method. The dynamic behavior of rockfill materials was modeled with a viscoelastic model and an empirical permanent strain model. The relevant parameters were obtained either by back analysis using the field observations or by reference to parameters of similar rockfill materials. The acceleration responses of the dam, the distribution of earthquake-induced settlement, and the gap propagation under the concrete slabs caused by the settlement of the dam were analyzed and compared with site investigations or relevant studies. The mechanism of failure of horizontal construction joints was also analyzed based on numerical results and site observations. Numerical results show that the input accelerations were considerably amplified near the top of the dam, and the strong shaking resulted in considerable settlement of the rockfill materials, with a maximum value exceeding 90 cm at the crest. As a result of the settlement of rockfill materials, the third-stage concrete slabs were separated from the cushion layer. The rotation of the cantilever slabs about the contacting regions, under the combined action of gravity and seismic inertial forces, led to the failure of the construction joints and tensile cracks appeared above the construction joints. The effectiveness and limitations of the so-called equivalent linear method are also discussed.

Background/Objectives: Our aim was to investigate the calcium content of different gallstone compositions and the pathogenic mechanisms of calcium carbonate gallstones. Methods: Between August 2001 and July 2007, gallstones from 481 patients, including 68 calcium carbonate gallstones, were analyzed for total calcium content. Gallbladder bile samples from 33 cases and six controls were analyzed for pH, carbonate anion level, free-ionized calcium concentration and saturation index for calcium carbonate. Results: Total calcium content averaged 75.6 %, 11.8 %, and 4.2 % for calcium carbonate, calcium bilirubinate and cholesterol gallstones. In 29.4 % of patients, chronic and/or intermittent cystic duct obstructions were caused by polypoid lesions in the neck region and 70.6 % were caused by stones. A total of 82 % of patients had chronic low-grade inflammation of the gallbladder wall and 18.0 % had acute inflammatory exacerbations. In the bile, we found the mean pH, mean carbonate anion, free-ionized calcium concentrations, and mean saturation index for calcium carbonate to be elevated in comparison to controls. Conclusion: From our study, we found chronic and/or intermittent cystic duct obstructions and low-grade GB wall inflammation lead to GB epithelium hydrogen secretion dysfunction. Increased calcium ion efflux into the GB lumen combined with increased carbonate anion presence increases SI_CaCO3 from 1 to 22.4. Thus, in an alkaline milieu with pH 7.8, calcium carbonate begins to aggregate and precipitate.

Acute respiratory distress syndrome (ARDS) is a major cause of mortality and morbidity in adult intensive care units. The relationship between the timing of ARDS onset and outcome is not well known. The objective of this study was to investigate the outcome of patients with late-onset ARDS during hospitalization. Methods: We prospectively enrolled patients who were intubated and fulfilled ARDS criteria in medical and surgical intensive care units in a tertiary referral medical center from December 1, 2004 to May 31, 2006. Those who developed ARDS more than 48 hours after hospital admission were categorized as late- onset ARDS; otherwise, they were defined as early-onset ARDS. We assessed the risk factors for hospital mortality using multivariate analysis and 90-day survival using Kaplan–Meier analysis between early- and late-onset ARDS, and between direct and indirect ARDS. Results: A total of 172 patients were included in the study. Overall mortality rate was 70%. Late-onset ARDS [odds ratio (OR): 3.06; 95% confidence interval (CI): 1.41 to 6.63; p = 0.005] and initial shock (OR: 8.20; 95% CI: 3.39–19.79; p < 0.001) were the independent risk factors for hospital mortality. Patients with late-onset ARDS had higher hospital mortality rate (83% vs. 60%; p = 0.002), longer duration of mechanical ventilation (27.0 ± 23.4 vs.14.6 ± 11.5 days; p < 0.001) and length of intensive care unit stay (25.5 ± 20.6 vs. 15.6 ± 13.6 days; p < 0.001) than patients with early-onset ARDS. The 90-day survival showed that both early-onset ARDS and direct ARDS were associated with better survival. Conclusion: Patients with late-onset ARDS are associated with poor prognosis and should be managed as high-risk patients.

Background & Aims: Intrahepatic cholangiocarcinoma (iCCA) is the second most common primary liver cancer and is highly lethal. Clonorchis sinensis (C. sinensis) infection is an important risk factor for iCCA. Here we investigated the clinical impact and underlying molecular characteristics of C. sinensis infection-related iCCA., Methods: We performed single-cell RNA sequencing, whole-exome sequencing, RNA sequencing, metabolomics and spatial transcriptomics in 251 patients with iCCA from three medical centers. Alterations in metabolism and the immune microenvironment of C. sinensis-related iCCAs were validated through an in vitro co-culture system and in a mouse model of iCCA., Results: We revealed that C. sinensis infection was significantly associated with iCCA patients' overall survival and response to immunotherapy. Fatty acid biosynthesis and the expression of fatty acid synthase (FASN), a key enzyme catalyzing long-chain fatty acid synthesis, were significantly enriched in C. sinensis-related iCCAs. iCCA cell lines treated with excretory/secretory products of C. sinensis displayed elevated FASN and free fatty acids. The metabolic alteration of tumor cells was closely correlated with the enrichment of tumor-associated macrophage (TAM)-like macrophages and the impaired function of T cells, which led to formation of an immunosuppressive microenvironment and tumor progression. Spatial transcriptomics analysis revealed that malignant cells were in closer juxtaposition with TAM-like macrophages in C. sinensis-related iCCAs than non-C. sinensis-related iCCAs. Importantly, treatment with a FASN inhibitor significantly reversed the immunosuppressive microenvironment and enhanced anti-PD-1 efficacy in iCCA mouse models treated with excretory/secretory products from C. sinensis., Conclusions: We provide novel insights into metabolic alterations and the immune microenvironment in C. sinensis infection-related iCCAs. We also demonstrate that the combination of a FASN inhibitor with immunotherapy could be a promising strategy for the treatment of C. sinensis-related iCCAs., Impact and Implications: Clonorchis sinensis (C. sinensis)-infected patients with intrahepatic cholangiocarcinoma (iCCA) have a worse prognosis and response to immunotherapy than non-C. sinensis-infected patients with iCCA. The underlying molecular characteristics of C. sinensis infection-related iCCAs remain unclear. Herein, we demonstrate that upregulation of FASN (fatty acid synthase) and free fatty acids in C. sinensis-related iCCAs leads to formation of an immunosuppressive microenvironment and tumor progression. Thus, administration of FASN inhibitors could significantly reverse the immunosuppressive microenvironment and further enhance the efficacy of anti-PD-1 against C. sinensis-related iCCAs., (Copyright © 2024 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Introduction: Secondary spontaneous pneumothorax (SSP) is often linked to chronic obstructive pulmonary disease (COPD). The frequency of SSP occurrence in COPD patients varies among different research findings. SSPs are more commonly found in the elderly population diagnosed with COPD. Previous studies have reported a pneumothorax rate of 26 per 100,000 COPD patients. There is, however, a notable lack of detailed epidemiological information regarding SSP in Asia. Our study focused on determining the occurrence rate of SSP among COPD patients in Taiwan using an extensive national database. Additionally, this study aimed to identify comorbidities associated with SSP in this patient group., Methods: In this study, we used the Longitudinal Health Insurance Database, which contains records of 2 million people who were randomly chosen from among the beneficiaries of the Taiwan National Health Insurance program. The dataset includes information from 2005 to the end of 2017. Our focus was on individuals diagnosed with COPD, identified through ICD-9-CM codes in at least one hospital admission or two outpatient services, with the COPD diagnosis date as the index date. The exclusion criteria included individuals younger than 40 years, those with incomplete records, or those with a previous diagnosis of pneumothorax before the index date. We conducted a matched comparison by pairing COPD patients with control subjects of similar age, sex, and comorbidities using propensity score matching. The follow-up for all participants started from their index date and continued until they developed pneumothorax, reached the study's end, withdrew from the insurance program, or passed away. The primary objective was to evaluate and compare the incidence of pneumothorax between COPD patients and matched controls., Results: We enrolled 65,063 patients who were diagnosed with COPD. Their mean age (±SD) was 66.28 (±12.99) years, and approximately 60 % were male. During the follow-up period, pneumothorax occurred in 607 patients, equivalent to 9.3 % of the cohort. The incidence rate of SSP in COPD patients was 12.10 per 10,000 person-years, whereas it was 6.68 per 10,000 person-years in those without COPD. Furthermore, COPD patients with comorbidities such as atrial fibrillation, congestive heart failure, coronary artery disease, diabetes mellitus, hypertension, and cancer exhibited an increased incidence of SSP compared to COPD patients without such comorbidities. This was observed after conducting a multivariable Cox regression analysis adjusted for age, sex, and other comorbidities., Conclusion: Our study revealed an elevated risk of SSP in patients with COPD. It has also been suggested that COPD patients with comorbidities, such as atrial fibrillation, congestive heart failure, coronary artery disease, diabetes mellitus, hypertension, and cancer, have an increased risk of developing SSP., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. Kuang-Ming Liao reports article publishing charges was provided by Chi Mei Medical Center, Chiali. Kuang-Ming Liao reports a relationship with Chi Mei Medical Center, Chiali that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

To deeply explore the intervention effects of ischelium on the cognitive memory decline in naturally aging mice and its potential mechanisms, we randomly divided mice into four groups: young control group (C), elderly group (M), ischelium low-dose group (L), and ischelium high-dose group (H). The experiment lasted for 12 weeks. We employed the Y-maze test, open field test, and conditioned fear test to evaluate the memory functions of each group. Through HE staining and electron microscopy, we observed morphological changes in the mouse hippocampus. RT-PCR was used to detect changes in the expression of factors related to cognitive function in the hippocampus of elderly mice. We analyzed the changes in the Nrf2/HO-1 pathway and the inflammatory factors IL-1β and TNF-α using elisa. Additionally, we examined the enzymatic activities of SOD, CAT, GSH-Px, and MDA in the hippocampus and analyzed the compositional changes of gut microbiota in mice using 16S technology. Our results indicate that ischelium effectively ameliorates cognitive impairments in elderly mice., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: We investigated the impacts of the standard treatment durations of and adherence to standard anti-tuberculous therapy (ATT) on recurrence after the successful completion of tuberculosis (TB) treatment., Methods: We recruited patients with TB who had received treatment for six or nine months from the 2008-2017 databases of the Taiwanese National Health Insurance Research Database. Treatment duration and adherence to standard ATT were analyzed for their impacts on recurrence within two years. Complete adherence to standard ATT was defined as daily use of ethambutol, isoniazid, pyrazinamide, and rifampin for the first two months, and daily use of isoniazid and rifampin for the first six months., Results: A total of 33,298 TB patients with new-onset TB were identified and classified into two groups by treatment duration: six months (n = 25,849, 77.63%) and nine months (n = 7449). Sex and age distributions varied between the groups. Treatment duration did not affect TB recurrence within two years (adjusted hazard ratio (AHR): 1.18, 95% confidence interval (C.I.) [0.96-1.44], p = 0.1156). Multivariable logistic regression showed that incomplete adherence to standard anti-tuberculous therapy (80-89% and 90-99% standard anti-TB therapy, AHR: 1.57, 95% C.I. [1.26-1.95], and 1.63, 95% C.I. [1.26-2.06], respectively, p < 0.0001) increased TB recurrence. In addition, male sex, older age, and comorbidity with diabetes mellitus or chronic obstructive pulmonary disease were independent risk factors for TB recurrence within two years., Conclusions: TB recurrence was 1.54% within two years under a DOT era. TB treatment durations of six or nine months did not affect TB recurrence within two years after completion of TB treatment, but incomplete adherence to standard anti-tuberculous therapy might increase the TB recurrence rate., Competing Interests: Declaration of Competing Interest All authors have no competing interests to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background & Aims: RNA N 6 -methyladenosine (m 6 A) reader protein YTHDF1 has been implicated in cancer; however, its role in hepatocellular carcinoma (HCC), especially in non-alcoholic steatohepatitis-associated HCC (NASH-HCC), remains unknown. Here, we investigated the functional role of YTHDF1 in NASH-HCC and its interplay with the tumor immune microenvironment., Methods: Hepatocyte-specific Ythdf1-overexpressing mice were subjected to a NASH-HCC-inducing diet. Tumor-infiltrating immune cells were profiled with single-cell RNA-sequencing, flow cytometry, and immunostaining. The molecular target of YTHDF1 was elucidated with RNA-sequencing, m 6 A-sequencing, YTHDF1 RNA immunoprecipitation-sequencing, proteomics, and ribosome-profiling. Ythdf1 in NASH-HCC models was targeted by lipid nanoparticle (LNP)-encapsulated small-interfering Ythdf1., Results: YTHDF1 is overexpressed in tumor tissues compared to adjacent peri-tumor tissues from patients with NASH-HCC. Liver-specific Ythdf1 overexpression drives tumorigenesis in dietary models of spontaneous NASH-HCC. Single-cell RNA-sequencing and flow cytometry revealed that Ythdf1 induced accumulation of myeloid-derived suppressor cells (MDSCs) and suppressed cytotoxic CD8 + T-cell function. Mechanistically, Ythdf1 expression in NASH-HCC cells induced the secretion of IL-6, which mediated MDSC recruitment and activation, leading to CD8 + T-cell dysfunction. EZH2 mRNA was identified as a key YTHDF1 target. YTHDF1 binds to m 6 A-modified EZH2 mRNA and promotes EZH2 translation. EZH2 in turn increased expression and secretion of IL-6. Ythdf1 knockout synergized with anti-PD-1 treatment to suppress tumor growth in NASH-HCC allografts. Furthermore, therapeutic targeting of Ythdf1 using LNP-encapsulated small-interfering RNA significantly increased the efficacy of anti-PD-1 blockade in NASH-HCC allografts., Conclusions: We identified that YTHDF1 promotes NASH-HCC tumorigenesis via EZH2-IL-6 signaling, which recruits and activates MDSCs to cause cytotoxic CD8 + T-cell dysfunction. YTHDF1 may be a novel therapeutic target to improve responses to anti-PD-1 immunotherapy in NASH-HCC., Impact and Implications: YTHDF1, a N 6 -methyladenosine reader, is upregulated in patients with non-alcoholic steatohepatitis (NASH)-associated hepatocellular carcinoma (HCC); however, its role in modulating the tumor immune microenvironment in NASH-HCC remains unclear. Here, we show that Ythdf1 mediates immunosuppression in NASH-HCC and that targeting YTHDF1 in combination with immune checkpoint blockade elicits robust antitumor immune responses. Our findings suggest novel therapeutic targets for potentiating the efficacy of immune checkpoint blockade in NASH-HCC and provide the rationale for developing YTHDF1 inhibitors for the treatment of NASH-HCC., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: Type 2 diabetes mellitus (DM) is a risk factor for mycobacterial pulmonary infections (MPI), including tuberculosis (TB) and nontuberculous mycobacterial lung disease (NTM-LD). Dipeptidyl peptidase IV inhibitor (DPP4i), a common DM medication, has an immune-modulation effect that raises concerns about developing MPI. However, there is scarce research on the topic., Methods: This retrospective study was conducted in a tertiary-referral center in Taiwan from 2009 to 2016. Patients with type 2 DM who were receiving any DM medication were enrolled. TB and NTM-LD were defined by microbiological criteria. We analyzed the risk of MPI in DPP4i users using Cox proportional hazard regression with adjusted inverse probability of treatment weighting., Results: A total of 9963 patients were included. Among them, 3931 were classified as DPP4i users, and 6032 patients were DPP4i nonusers. DPP4i users had no increase in incidences of MPI (604 vs. 768 per 100,000 person-years, p = 0.776), NTM-LD (174 vs. 255 per 100,000 person-years, p = 0.228), and TB (542 vs. 449 per 100,000 person-years, p = 0.663) relative to those of DPP4i nonusers. After adjustment, the adjusted hazard ratios for MPI (aHR: 1.07, 95% CI: 0.79-1.45), TB (aHR: 1.15, 95% CI: 0.81-1.64) and NTM-LD (aHR: 0.85, 95% CI: 0.49-1.47) were not significantly increased relative to those of nonusers. The subgroup analysis also showed that DPP4i use did not increase the risk of MPI in different DM severities and comorbidities., Conclusions: According to our large cohort study, DPP4i use is safe for patients with type 2 DM and might not increase the risk of MPI., Competing Interests: Declaration of Competing Interest No conflict of interest to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background & Aims: The immune landscape of hepatocellular carcinoma (HCC) following transarterial chemoembolisation (TACE) remains to be clarified. This study aimed to characterise the immune landscape following TACE and the underlying mechanism of HCC progression., Methods: Tumour samples from five patients with treatment-naive HCC and five patients who received TACE therapy were collected and subjected to single-cell RNA sequencing. Another 22 paired samples were validated using immunofluorescence staining and flow cytometry. To clarify the underlying mechanisms, in vitro co-culture experiments and two types of TREM2-KO/WT mouse models, namely, an HCC cell orthotopic injection model and a spontaneous HCC model, were used., Results: A reduced number of CD8 + T cells and an increased number of tumour-associated macrophages (TAMs) were observed in the post-TACE microenvironment. TACE therapy reduced the cluster CD8&#95;C4, which was highly enriched with tumour-specific CD8 + T cells of pre-exhausted phenotype. TREM2 was found to be highly expressed in TAMs following TACE, which was associated with a poor prognosis. TREM2 + TAMs secreted less CXCL9 but more galectin-1 than did TREM2 - TAMs. Galectin-1 promoted PD-L1 overexpression in vessel endothelial cells, impeding CD8 + T cell recruitment. TREM2 deficiency also increased CD8 + T cell infiltration, which inhibited tumour growth in both in vivo HCC models. More importantly, TREM2 deficiency enhanced the therapeutic effect of anti-PD-L1 blockade., Conclusions: This study shows that TREM2 + TAMs play an important role in suppressing CD8 + T cells. TREM2 deficiency increased the therapeutic effect of anti-PD-L1 blockade by enhancing antitumour activity of CD8 + T cells. These findings explain the reasons for recurrence and progression after TACE and provide a new target for HCC immunotherapy after TACE., Impact and Implications: Studying the immune landscape in post-TACE HCC is important to uncover the mechanisms of HCC progression. By using scRNA sequencing and functional assays, we discovered that both the number and function of CD8 + T cells are compromised, whereas the number of TREM2 + TAMs is increased in post-TACE HCC, correlating with worse prognosis. Moreover, TREM2 deficiency dramatically increases CD8 + T cell infiltration and augments the therapeutic efficacy of anti-PD-L1 blockade. Mechanistically, TREM2 + TAMs display lower CXCL9 and increased Gal-1 secretion than do TREM2 - TAMs, with Gal-1 mediating the overexpression of PD-L1 in vessel endothelial cells. These results suggest that TREM2 could be a novel immunotherapeutic target for patients treated with TACE in HCC. This provides an opportunity to break the plateau of limited therapeutic effect. This study has the value of understanding the tumour microenvironment of post-TACE HCC and thinking a new strategy of immunotherapy in the field of HCC. It is therefore of key impact for physicians, scientists and drug developers in the field of liver cancer and gastrointestinal oncology., (Copyright © 2023 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Background and Study Aims: Post-hepatectomy liver failure (PHLF) is the main cause of perioperative death after hepatocellular carcinoma (HCC) resection. PHLF occurrence is related to both the hepatectomy volume and the degree of cirrhosis. Accurate preoperative assessment of the degree of cirrhosis may aid in reducing the incidence of PHLF. Several studies have shown that the liver stiffness measurement (LSM) is well correlated with cirrhosis. This study explored the relationship between LSM and PHLF occurrence after radical HCC resection and the effect on long-term prognosis., Patients and Methods: We retrospectively analyzed the clinical data of 164 patients who underwent radical HCC resection at our center from January 2017 to January 2020. The related postoperative PHLF factors were analyzed. The LSM threshold in postoperative PHLF was calculated through receiver operating characteristic (ROC) curve analysis. Patients were grouped according to different LSM thresholds and survival analysis was performed., Results: Forty-six patients experienced PHLF, of whom 19, 21, and 6 were classified as grades A, B, and C, respectively. Multivariate analysis indicated that LSM was an independent risk factor for PHLF after HCC surgery (OR = 1.174, P < 0.000). LSM (OR = 1.219, P < 0.000) and intraoperative bleeding (OR = 1.001, P = 0.047) were risk factors for grade B-C PHLF. The LSM threshold that predicted PHLF occurrence was 17.9 kPa (AUC = 0.831, P < 0.000) and 24.5 kPa (AUC = 0.867, P < 0.000) for grade B-C PHLF. LSM was correlated with PHLF severity (r = 0.439, P < 0.001). The median survival times were 32 vs 26 months (P = 0.016) for patients with LSM ≤ 17.9 kPa vs those with LSM > 17.9 kPa and 28 vs 24 months (P = 0.004) for patients with LSM ≤ 24.5 kPa vs those with LSM > 24.5 kPa., Conclusion: LSM is related to PHLF occurrence in patients undergoing HCC resection; a higher LSM is associated with the occurrence of more severe PHLF after surgery. In addition, LSM may aid in predicting long-term survival after liver resection in patients with HCC., (Copyright © 2022 Pan-Arab Association of Gastroenterology. Published by Elsevier B.V. All rights reserved.)

Background: Lung cancer patients are high-risk for active tuberculosis (TB); however, fragility and drug-drug interaction might lead to TB treatment interruption. TB treatment incompletion occurrence and predictors among lung cancer patients remain unclear., Methods: We recruited lung cancer patients with new-onset TB from Taiwan Cancer Registry and Taiwanese National Health Insurance 2007-2015 databases. TB treatment incompletion was the identified primary outcome, and associated risk factors were analyzed., Results: A total of 1155 lung cancer patients with new-onset TB were identified and classified as treatment incompletion (n=706, 61.13%) or completion (n=449). Gender and age distribution was similar in both groups. Under multivariable logistic regression, advanced cancer (stage III and IV) and no first-line TB drugs use were independent factors for treatment incompletion; but older age was not significant. For patients surviving >1 year since TB diagnosis, independent factors for treatment incompletion included no first-line TB drugs use (except pyrazinamide) and absence of hypertension. Cancer stage had borderline significance., Conclusions: TB treatment incompletion occurred in 61.13% of lung cancer patients. Clinicians should carefully titrate anti-TB medications and monitor side effects in lung cancer patients, especially those with treatment incompletion risk factors, to avoid treatment interruption due to fragility and/or drug intolerance., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Coronavirus-caused pneumonia (COVID-19) broke out in December 2019. The virus soon proved to be extremely contagious and caused an international pandemic. Clinicians treating COVID-19 patients face considerable danger of occupational exposure because of the highly infectious nature of the virus, and precautions must be taken to prevent medical staff infections. This article lists important measures that may save the lives of patients and medical staff during the COVID-19 pandemic and help stop the transmission of COVID-19 on hospital grounds. The suggestions include establishing detailed infection control and prevention protocols in the operating room; expediting testing procedures and patient screening for COVID-19; using case-specific treatment planning for vascular patients with COVID-19, favoring minimally invasive methods; and establishing and reinforcing protective awareness of medical personnel., (Copyright © 2020 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Disuse osteoporosis (DOP) is a common complication resulting from the lack of or disuse of mechanical loading and has been unsatisfactorily treated. We hypothesized that exosomes derived from human umbilical cord mesenchymal stem cells (HUCMSCs) could reduce bone marrow mesenchymal stem cell (BMSC) apoptosis in rat DOP via the miR-1263/Mob1/Hippo signaling pathway. To evaluate the function of exosomes derived from HUCMSCs (HUCMSC-Exos) in DOP, hind limb unloading (HLU)-induced DOP rat models were prepared. In vitro, the proliferation of BMSCs were evaluated using CCK-8 assays. Further, the apoptosis of BMSCs were evaluated using annexin V-FITC assay and Western blots. In vivo, the protective effects of HUCMSC-Exos were evaluated using HE staining and microCT analysis. The underlying molecular mechanism of exosome action on BMSC apoptosis through the miR-1263/Mob1/Hippo pathway was also investigated by high-throughput RNA sequencing, luciferase reporter assays, RNA-pull down assays and Western blots. The RNA-seq and q-PCR results showed that the level of miR-1263 was most abundant among differentially expressed microRNAs. Exosomal miR-1263 could bind to the 3'untranslated region (3' UTR) of Mob1 and exert its function by directly targeting Mob1 in recipient cells. The inhibition of Mob1 could activate YAP expression. Hippo inhibition reversed the in vitro HLU-induced apoptotic effect on BMSCs. The microCT and HE staining results indicated that HUCMSC-Exos ameliorated DOP in vivo. Exosomes derived from HUCMSCs are effective at inhibiting BMSC apoptosis and preventing rat DOP. This mechanism is mediated by the miR-1263/Mob1/Hippo signaling pathway., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Purpose: Glucocorticoid-induced osteonecrosis of the femoral head (GIONFH) is a common disease following long-term use or large doses of glucocorticoids. The pathogenesis of GIONFH remains controversial, and abnormal bone metabolism caused by glucocorticoids(GCs) may be one of the important factors. Due to its positive effect on bone remodeling, naringin shows potential therapeutic effects in bone metabolism-related diseases. In this study, we hypothesized that naringin regulated bone metabolism in rat GIONFH via the Akt/Bad signal cascades., Methods: In vitro, a dexamethasone (Dex)- or naringin-treated cell model was used to evaluate the function of naringin. In vivo, methylprednisolone (MPS)-treated rat model was used to evaluate the function of naringin in GIONFH. In vitro, Cell Counting Kit-8 (CCK-8) and Edu staining was used to evaluate the proliferation of osteocytes, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, Annexin V-FITC-PI, and western blotting were used to evaluate the apoptosis of osteocytes. We also verified the effects of naringin on osteogenesis and osteoclastogenesis. In vivo, we used micro-CT (computed tomography), histological, and immunohistochemical analysis to evaluate the effect of naringin. Moreover, the mechanism of naringin regulating the bone metabolism through the Akt/Bad pathway was also investigated using bioinformatics analysis and western blotting., Results: The results of in vitro study showed that Akt activated by naringin promoted osteogenesis and osteocyte proliferation; in addition, osteocyte apoptosis and osteoclastogenesis was inhibited by Akt activation and Bad suppression. According to the in vivo study, naringin prevented GIONFH in a rat model as shown by micro-CT scanning and histological and immunohistochemical analysis., Conclusions: Therefore, we concluded that naringin is an effective compound for promoting bone repair and preventing bone loss in rats with GIONFH through Akt/Bad signal cascades., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: large doses of glucocorticoids (GCs) are the most common cause of glucocorticoid-induced osteonecrosis of femoral head (GIONFH). Although awareness of GIONFH among patients with GCs history has increased over recent years, several studies indicate that its mechanism remains unclear., Methods: To evaluate the function of circUSP45 in GIONFH, femoral heads in GIONFH patients or femoral heads in fracture patients were collected. In vitro, RT-PCR, FISH, RNA pull down and Western blotting assay were used to evaluate the function of circUSP45. In addition, we also verified the effects of circUSP45 on osteogenesis using alizarin red staining. In vivo, we used HE staining and microCT analysis to evaluate the bone mass. Moreover, the mechanism of circUSP45 regulating osteogenesis through the miR-127-5p/PTEN/AKT pathway was also investigated., Results: The results showed that expression of circUSP45 increased in GIONFH patients. The overexpression of circUSP45 decreases osteogenic gene expression and inhibits the proliferation of BMSCs. Furthermore, circUSP45 was located mainly in the cytoplasm and directly interacted with miR-127-5p. MiR-127-5p acts with its targets PTEN to regulate the osteogenesis. MicroCT and HE staining verify the function of circUSP45 in GIONFH rat model., Conclusion: CircUSP45 decreases osteogenesis in bone GIONFH by sponging miR-127-5p through PTEN/AKT signal pathway., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Aims: To establish multiparametric radiomics of rectal tumor for the preoperative prediction of lymph node (LN) metastasis., Materials and Methods: This prospective study consisted of 115 consecutive patients with rectal carcinoma between April 2015 and April 2017. The multiparametric radiomics scores were extracted from the endorectal ultrasound (ERUS), computed tomography (CT) and shear-wave elastography (SWE) features of the rectal tumor, LN, and peripheral tissues. The three radiomics scores were generated. Further validation as an independent predictor was performed using multivariate logistic regression together with clinical data, and a nomogram was subsequently developed. The predictive performance of the multiparametric radiomics nomogram was compared with that of conventional radiomics., Key Findings: All three scores (ERUS, CT, and SWE) were significantly higher in patients with LN metastasis than in patients with negative LN metastasis (all P < 0.05) in both training and validation set. Multivariate analysis indicated that CT and SWE scores were independent risk variables (odds ratio, OR = 6.764 and 5.482, respectively). In validation cohort, the multiparametric radiomics nomogram showed the highest predictive accuracy for LN metastasis, with a concordance index (C-index) of 0.857 compared with the conventional radiomics nomogram (C-index, 0.703, P = 0.100), resulting in a significantly improved net reclassification index (NRI) (P < 0.05) and integrated discriminatory improvement (IDI) (P = 0.002). Decision curve analysis showed that the multiparametric radiomics nomogram had a higher overall net benefit., Significance: Multiparametric radiomics of rectal cancer, which captures blood supply and stiffness phenotypes, is a useful tool for predicting LN metastasis preoperatively and has marked discrimination accuracy compared to conventional radiomics., (Copyright © 2018. Published by Elsevier Inc.)

Herein, we describe an improved procedure for the green synthesis of chondroitin sulfate stabilized silver nanoparticles (ChS-AgNPs). Glucose was used as a reducing agent under alkaline conditions to obtain a small particle size (<10 nm), and the reduction was complete within one hour at room temperature. The concentration of NaOH affected the reaction rate, formation yield, and particle size of ChS-AgNPs. The formation of AgNPs was confirmed using UV-vis, TEM, XRD, and XPS. ChS-AgNPs showed excellent catalytic activities in the reduction of 4-nitrophenol by NaBH 4 , and the reaction rate increased linearly with increasing catalyst amounts. The antimicrobial activities of ChS-AgNPs against A. baumannii (including multidrug-resistant strains), E. coli, P. aeruginosa, and S. aureus were evaluated using the broth microdilution method. Finally, from the morphological observations and cell cycle analysis of L929 cells, we found that ChS-AgNPs exhibited antimicrobial and biocompatible activities., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Stress-induced phosphoprotein 1 (STIP1) is an adaptor protein that bridges between HSP70 and HSP90 folding and a secretory protein which regulates malignant cell growth. However, the role of STIP1 in hepatocellular carcinoma (HCC) remains unknown. Here, we found high expression of STIP1 in tumors was associated with worse overall survival (41.3 vs 62.7 months, P < 0.001) in 231 HCC patients. STIP1 was overexpressed in HCC tissues compared to adjacent non-tumor liver tissue (64.9% vs 4.0% P < 0.001), and serum STIP1 levels of HCC patients were elevated compared to healthy controls (P < 0.001). Mechanistically, STIP1 promoted HCC growth through PI3K-AKT-dependent anti-apoptotic pathway. STIP1 mediated cell growth in an autocrine fashion, which could be suppressed either by neutralizing extracellular STIP1 or by knocking down intracellular STIP1. In xenograft mouse model, knockdown of STIP1 significantly reduced tumor growth (P < 0.001). In conclusion, STIP1 is upregulated in HCC and associated with poor clinical prognosis. Blocking STIP1 activity suppresses HCC cell growth, providing the rationale for STIP1 as a potential therapeutic target in HCC., (Copyright © 2017. Published by Elsevier Inc.)

The low-density lipoprotein receptor-related protein 1 (LRP1) gene is associated with increased levels of plasma factor VIII (FVIII). We aimed to explore eight functional genetic LRP1 variants for their potential roles in regulating FVIII levels and acute ischemic stroke (AIS). This genetic association study enrolled 192 patients with AIS and 134 controls. There were no significant differences in the genetic frequency of the eight functional single-nucleotide polymorphisms (SNPs) between the control and AIS groups. However, while analyzing the association between the eight SNPs and plasma FVIII levels, subjects with T/T genotype of rs1800137 (vs. CC+CT) were found to be associated with higher FVIII levels (23.5IU/dL; 95% confidence interval, 7.4-39.5IU/dL; P=0.0044) after adjusting for age, gender, estimated glomerular filtration rate, O blood type, inflammatory state, and body mass index. An analysis of the mRNA stability and abundance was designed and performed using minigene system transfected into HepG2 cells to assess the possible differences in mRNA stabilities between rs1800137 CC (rs1800137C) and TT (rs1800137T) genotypes. Site-directed mutagenesis revealed that rs1800137T accounts for the observed decrease in mRNA stability. The SNP rs1800137, located in exon 8, has been identified as an exon-splicing enhancer in silico. However, alternative splicing of LRP1 without inclusion of exon 8 was not identified. In transfected HepG2 cells, cycloheximide slowed down the degradation of the rs1800137T-containing minigene. These results demonstrate that synonymous SNP rs1800137 can lead to increased plasma FVIII levels due to decreased mRNA stability via translation-dependent mRNA degradation associated with codon optimality., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Hilar cholangiocarcinoma (HCCA) is a devastating malignancy arising from the bifurcation of the hepatic duct, whether combined vascular resection benefits HCCA patients is controversial. This study was undertaken to assess the effect of combined vascular resection in HCCA patients and to analyze the prognostic factors., Methods: Clinical data of 154 HCCA patients who had been treated from January 2005 to December 2012 were retrospectively analyzed. The patients were divided into three groups based on vascular resection: those without vascular resection; those with portal vein resection alone and those with hepatic artery resection. The survival and complication rates were compared among the three groups. Multivariate analysis was made to determine prognostic factors., Results: No significant differences were found in survival and complication rates among the three groups (P>0.05). Multivariate analysis showed that 3 factors were related to survival: lymph node metastasis, tumor size (>2.5 cm), and positive resection margin., Conclusions: Vascular resection improved the survival rate of patients with HCCA involving the hepatic artery or portal vein. Lymph node metastasis, tumor size (>2.5 cm) and positive resection margin were poor prognostic factors in patients with HCCA.

Background/objectives: Our aim was to investigate the calcium content of different gallstone compositions and the pathogenic mechanisms of calcium carbonate gallstones., Methods: Between August 2001 and July 2007, gallstones from 481 patients, including 68 calcium carbonate gallstones, were analyzed for total calcium content. Gallbladder bile samples from 33 cases and six controls were analyzed for pH, carbonate anion level, free-ionized calcium concentration and saturation index for calcium carbonate., Results: Total calcium content averaged 75.6 %, 11.8 %, and 4.2 % for calcium carbonate, calcium bilirubinate and cholesterol gallstones. In 29.4 % of patients, chronic and/or intermittent cystic duct obstructions were caused by polypoid lesions in the neck region and 70.6 % were caused by stones. A total of 82 % of patients had chronic low-grade inflammation of the gallbladder wall and 18.0 % had acute inflammatory exacerbations. In the bile, we found the mean pH, mean carbonate anion, free-ionized calcium concentrations, and mean saturation index for calcium carbonate to be elevated in comparison to controls., Conclusion: From our study, we found chronic and/or intermittent cystic duct obstructions and low-grade GB wall inflammation lead to GB epithelium hydrogen secretion dysfunction. Increased calcium ion efflux into the GB lumen combined with increased carbonate anion presence increases SI&#95;CaCO(3) from 1 to 22.4. Thus, in an alkaline milieu with pH 7.8, calcium carbonate begins to aggregate and precipitate., (Copyright © 2012. Published by Elsevier B.V.)

In this study, the potential of chondroitin sulfate (ChS)-chitosan (CS) nanoparticles (NPs) for the delivery of proteins was investigated. ChS-CS NPs were prepared by ionic cross-linking of CS solution with ChS. The aggregation line, particle size and zeta potential were investigated as a function of the pH, weight ratio and concentration. The water content and formation yield of the NPs were measured by gravimetry. Results indicated that ChS-CS NPs showed a higher degree of ionic cross-linking and formation yield than sodium tripolyphosphate-CS NPs. Fluorescein isothiocyanate conjugate bovine serum albumin (FITC-BSA), a model protein drug, was incorporated into the ChS-CS NPs. The encapsulation efficiency was obviously increased with the increase in initial FITC-BSA concentration and was as high as 90%. In vitro release studies of ChS-CS NPs showed a small burst effect following a continued and controlled release. Cytotoxicity tests with Caco-2 cells showed no toxic effects of ChS-CS NPs. The ex vivo cellular uptake studies using Caco-2 and HEK-293 cells indicated that NPs were found to be endocytosed into the cells. In conclusion, ChS-CS NPs are a potential new delivery system for the transport of hydrophilic compounds such as proteins., (Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The muscleblind-like (MBNL) proteins are tissue-specific alternative splicing regulators. Dysfunction of MBNL has been implicated in the pathogenesis of expanded CUG repeats-associated myotonic dystrophy (DM). In this study, we describe the identification and functional characterization of a Caenorhabditis elegans muscleblind (CeMbl) gene. CeMbl is a single gene alternatively spliced to generate two isoforms (CeMBL-A and CeMBL-B). It displays a high homology with human MBNL1 in the C3H1 zinc finger domains. CeMbl transcripts are detected in larval and adult stages. However, inactivation of CeMbl by RNA-mediated interference results in muscle phenotype only at adulthood. Immunofluorescence staining using anti-vinculin antibody reveals that the organization of dense body is disrupted in affected worms. Our results demonstrate a growth-dependent requirement of CeMbl on muscle structure and function. They also provide a possible molecular basis for the developmentally regulated toxicity of expanded CUG repeats.

In this study, we investigated the effect of NO donor, diethylamine/nitric oxide (DEA/NO), on the electrophysiological behavior of human skeletal muscle chloride channel (CLCN1). The wild-type and variants of CLCN1, including one polymorphism (P727L) and four mutants (T631I, D644G, G482R, and S471F), were expressed in Xenopus oocytes and the ionic current was measured by two-electrode voltage-clamp method. Our results revealed that there is no significant difference in the current-voltage relationships and half-voltage values of open probability between wild-type and variants of CLCN1 except for G482R. Application of the DEA-NO (0.1mM) significantly increases the channel conductance of wild-type, T631I, D644G, and S471F, but not P727L. This indicates that P727L polymorphism causes loss of sensitivity of CLCN1 to the DEA/NO treatment, which could be due to a conformational change caused by proline substitution. The data suggest that the polymorphic changes may affect the function of CLCN1 in response to the treatment of chemical compounds.

Expansion of CTG repeat within the 3'-untranslated region of the DMPK gene causes the most common neuromuscular disorder, myotonic dystrophy type 1 (DM1), through a RNA trans-dominant mechanism. Here, we explore Caenorhabditis elegans as a model system to investigate the repeat size-dependent toxic effect by expression of green fluorescent protein (GFP) transcripts with various lengths of untranslatable CUG repeats (CUG5, CUG30, CUG83, CUG125, and CUG213) in body wall muscles. CUG213 animals died during embryogenesis or showed retarded growth at larval stages due to defective muscle development. CUG125 animals, although can produce offspring, exhibited uncoordinated muscle function, deviated electropharyngeogram, and an age-dependent abnormality in muscle structure. Most CUG83 animals had normal muscle structure and function as those expressing 30 and shorter repeats. Our results demonstrate for the first time that the in vivo toxicity of CUG repeats is repeat length- and growth-regulated and suggest that expanded CUG repeats are sufficient to cause congenital-like phenotypes in living organisms.

Mutations in the CLCN1 gene frequently associate with myotonia congenita (MC). We have recently reported several CLCN1 mutants in Taiwanese patients. To further elucidate the correlation between the genotypes and phenotypes, in this study, we used Xenopus oocyte as a system to investigate the functional effects of these mutants. The fs793X and G482R mutants, which were suggested to have a dual inheritance pattern, were found to cause a functional loss of CLCN1 channels. While co-expression of fs793X and wild-type (WT) showed a reduction of chloride conductance by about half of WT channels, the activation curve of voltage-dependence was not shifted. A compound heterozygous mutant, P575S/D644G, was found in a patient. When both mutants were co-expressed in oocytes, they caused a shift of the voltage-dependence of activation curve to more positive values than individual mutant. This indicates that both P575S and D644G mutants may contribute cooperatively to change the gating property of CLCN1 channel. Interestingly, the S471F mutant did not cause significant alternation of functional properties. Consistent with the fact that T631I mutant was found in three asymptomatic individuals, the electrophysiological parameters of T631I were similar to those of WT CLCN1 channels, suggesting that T631I is a neutral mutation. These results further clarify the correlation between the mutations and their functional implications of CLCN1 channels.

We present the clinical manifestations, brain magnetic resonance images (MRI), and genetic analysis of a family with 2 siblings with congenital myotonic dystrophy type 1 (DM1) and 4 patients with classic DM1. These 2 patients with congenital DM1 had severe mental retardation and a characteristic feature of hyperintensity of white matter at the posterior-superior trigone (HWMPST), in addition to ventricular dilatation in T2-weighted images (T2WI) of brain MRI. In 2 of the 4 classic DM1 patients, brain T2WI MRI showed hyperintensity lesions in the bilateral frontal and/or temporal regions, which were absent in congenital DM1. In conclusion, we suggest that the HWMPST in brain MRI is a characteristic finding in congenital DM1, and that the severe cognitive impairments are not only attributable to the subcortical white matter lesions. In congenital DM1, the cognitive function is a diffuse impairment, which is different from that in classic DM1.

p73 is one of the p53 family members which are transcription factors involved in the regulation of cell proliferation, apoptosis, and differentiation. In this study, we cloned the p73 cDNA from zebrafish ovary RNA. The consensus open reading frame (1923bp) encodes a polypeptide of 640 amino acids which shares 70-95% identity to the p73 of other vertebrates. Expression of zebrafish p73 mRNA is restricted to tissues such as skin, fin, brain, ovary, and testis, in contrast to the ubiquitous expression of zebrafish p53 and p63. During embryonic development, p73 transcripts are detected from the zygote period to the early larva stage. Whole-mount in situ hybridization reveals that p73 expression is in the brain, including olfactory bulbs, telencephalon, and hypothalamus, as well as in the pharyngeal arches and the nose. Moreover, p73 protein is found in the ovary and testis sections by immunohistochemical staining.